Quinoline, isoquinoline and phthalazine derivatives as antagonists of the gonadotropin-releasing hormone

ABSTRACT

The invention relates to new quinoline, isoquinoline and phthalazine derivatives as antagonists of the gonadotropin-releasing hormone.

[0001] This application claims the benefit of the filing date of U.S.Provisional Application Serial No. 60/274,914 filed Mar. 12, 2001,incorporated in its entirety herein.

DESCRIPTION

[0002] The gonadotropin-releasing hormone (GnRH) is a hormone that issynthesized predominantly but not exclusively in mammals by nerve cellsof the hypothalamus, is transported via the portal vein to thehypophysis and is released in a regulated manner to the gonadotropiccells. By interaction with its receptor that has seven transmembranedomains, GnRH stimulates the production and the release of gonadotropichormones by means of the second messenger inositol-1,4,5-triphosphateand Ca²⁺ ions. The gonadotropin-luteinizing hormone (LH) that isreleased by GnRH and the follicle-stimulating hormone (FSH) stimulatethe production of sex steroids and the gamete maturation in both sexes.In addition to GnRH (also referred to as GrRH1), there are two otherforms of GnRH, namely GnRH2 and 3.

[0003] The GnRH receptor is used as a pharmacological target in a numberof diseases that are dependent on a functioning sex hormone production,for example prostate cancer, premenopausal breast cancer, endometriosisand uterine fibroids. In the case of these diseases, GnRH superagonistsor GnRH antagonists can be used successfully. In particular, the malebirth control in combination with a substitution dose of androgens formsa possible further indication.

[0004] An advantage of GnRH antagonists in comparison to superagonistsis their immediate effectiveness in the blocking of the gonadotropinsecretion. Superagonists initially produce an overstimulation of thehypophysis, which results in increased gonadotropin and sex steroidreleases.

[0005] This hormonal reaction is only completed after a certain delaybased on the desensitization and downward-adjustment of the GnRHreceptor concentrations. Therefore, GnRH superagonists, both alone andin combination with testosterone, may not be able to suppresseffectively sperm production in males and thus are not suitable for malebirth control. In contrast to this, peptide GNRH antagonists, especiallyin combination with a substitution dose of androgen, are able to bringabout a significant oligozoospermia in humans.

[0006] Peptide GnRH antagonists, however, have a number of drawbacks.They have a considerably lower effectiveness as superagonists andconsequently have to be administered at considerably higher dosages.Their oral bio-availability is also low, so that they have to beadministered by injection. Repeated injections lead in turn to areduction in compliance. Moreover, the synthesis of peptide GnRHantagonists in comparison to non-peptide compounds is costly andlabor-intensive.

[0007] Quinoline derivatives as non-peptide GnRH antagonists aredisclosed in, for example, WO97/14682. To date, however, it was notpossible to market any non-peptide GnRH antagonists.

[0008] The object on which this invention is based consisted inproviding new GnRH antagonists that are superior to the known peptidecompounds and represent an effective alternative to known non-peptidecompounds. The new GnRH antagonists are to have both high effectivenessand high oral bio-availability. In addition, they should be able to besynthesized simply and with as low costs as possible.

[0009] This object is achieved by compounds of general formula (1):

[0010] in which

[0011] R¹ (a) is an acyl group —CO—R11 or CN, whereby R11 is asaturated, unsaturated, cyclic and/or (hetero)aromatic organic radical,especially a straight or branched alkyl chain with 1-10 C atoms or aphenyl, furan or thiophene group that is optionally substituted by alkylgroups or halogen atoms,

[0012] (b) is a carboxylic acid ester group —CO—OR12 or a carboxylicacid amide group —CO—NR12R13 or a group —SO_(x)—R12 with X=0, 1 or 2 or—SO₂—NR12R13, whereby R12 is a saturated, unsaturated, cyclic and/or(hetero)aromatic organic radical, especially a straight or branchedalkyl chain with 1-10 C atoms, an aralkyl group with 7-20 C atoms,whereby the aryl radical optionally can be substituted by alkyl groupsor halogen atoms or is a phenyl radical that is optionally substitutedby alkyl groups or halogen atoms, and R13 can be a hydrogen atom or astraight or branched alkyl chain with 1-10 C atoms, or

[0013] (c) is the group -A—NR14—CO—NR15R16, in which A is an alkylenegroup with 1-4 C atoms, especially with 1 C atom, that is optionallysubstituted by a C₁-C₆ alkyl group, a carbonyl group, an oxygen atom orthe group —SO_(x)— with X=0, 1 or 2; R14 and R15, in each caseindependently are a hydrogen atom or a straight or branched alkyl chainwith 1-10 C atoms, and R16 is a straight or branched alkyl chain with1-10 C atoms, a cycloalkyl group with 3-10 C atoms, a cycloalkylalkylgroup with 7-20 C atoms, an aralkyl group with 7-20 C atoms, whereby thearyl radical optionally can be substituted by alkyl groups or halogenatoms, a phenyl group that is optionally substituted by alkyl groups orhalogen atoms or a heterocyclic ring that is optionally substituted byalkyl groups or halogen atoms,

[0014] R2 is a group —CH(R21)R22, whereby R21 is a hydrogen atom, aC₁-C₁₀-alkyl group or an optionally substituted phenyl ring and R22 isan optionally substituted phenyl ring or naphthyl ring, or a group—CH₂CH(R23)R24, with R23 and R24 in the meaning of an optionallysubstituted phenyl ring,

[0015] R3 and R4 in each case independently are a hydrogen atom or analkyl group with 1-10 C atoms, and R3 also can be a halogen atom,

[0016] R5 is a group that is linked via radical Z,

[0017] in which G is —C═C—, —C═N—, —N═C—, an oxygen or sulfur atom; Z isa direct bond, an oxygen atom or a sulfur atom, the group CH-R52 or—CHR52—CH—R53-, whereby R52 and R53, independently of one another, havethe meaning of a hydrogen atom or an alkyl group and n means numbers 1and 2, a —C≡C-triple bond or an E- or Z-configured group —CR52=CR53- orC═CR52R53, whereby R52 and R53, independently of one another, have themeaning of a hydrogen atom or an alkyl group, L is a CH₂ group or an NHgroup, Q is a carbonyl or —SO_(x) group, with X=0, 1 or 2, and R51 is anamino group that is optionally substituted by an alkyl group or astraight or branched alkyl group that is optionally substituted byhalogen atoms, hydroxyl or alkoxy groups, or a cycloalkyl group with 3-7ring members that is optionally substituted by halogen atoms, hydroxylor alkoxy groups,

[0018] R6 is the group CH₂—N(R61)R62, whereby R61, in each caseindependently, is a hydrogen atom or an alkyl group, and R62 is an alkylgroup or an optionally substituted aralkyl group or a heteroarylalkylgroup with 7-20 C atoms, and can mean

[0019] —W═X═Y— the groups

[0020]  in any orientation; also all stereoisomers of theabove-mentioned structures and salts thereof with physiologicallycompatible acids or bases.

[0021] In the compounds of formula (1), by way of example

[0022] R1 means:

[0023] A straight or branched alkyl chain: A methyl, ethyl, n-propyl,iso-propyl, n-, iso-, tert-butyl, n-pentyl, 2,2-dimethylpropyl or3-methylbutyl group; an n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decylgroup. The methyl or ethyl group is preferred.

[0024] A phenyl group that is optionally substituted by alkyl groups orhalogen atoms: A phenyl group; an o-, m-, p-methyl, ethyl, propyl, orisopropylphenyl group; a 2,3-, 2,4-, 2,5-, 2,6-, 3,4-, 3,5-dimethyl or-diethylphenyl group; an o-, m-, p-fluoro-, chloro-, bromo- oriodophenyl group; a 2,3-, 2,4-, 2,5-, 2,6-, 3,4-, 3,5-, difluoro-,dichloro-, dibromo- or diodophenyl group or a naphthyl group. A phenylgroup is preferred.

[0025] An optionally substituted furan or thiophene group: Anunsubstituted 2- or 3-thienyl group; or a 2- or 3-furyl group; or a3-methyl-, 3-ethyl-, 3-fluoro-, 3-chloro-, 3-bromo-, 3-iodo-2-furyl- or-2-thienyl group; a 4-methyl-, 4-ethyl-, 4-fluoro-, 4-chloro-, 4-bromo-,4-iodo-2-furyl- or 2-thienyl group; a 5-methyl-, 5-ethyl-, 5-fluoro-,5-chloro-, 5-bromo-, 5-iodo-2-furyl or -2-thienyl group; a 2-methyl-,2-ethyl-, 2-fluoro-, 2-chloro-, 2-bromo-, 2-iodo-3-furyl or -3-thienylgroup; a 4-methyl-, 4-ethyl-, 4-fluoro-, 4-chloro-, 4-bromo-,4-iodo-3-furyl- or -3-thienyl group; a 5-methyl-, 5-ethyl-, 5-fluoro-,5-chloro-, 5-bromo-, 5-iodo-3-furyl- or -3-thienyl group. Preferred is a2-thienyl or 2-furyl group.

[0026] An aralkyl group with 7-20 C atoms: A benzyl group; a1-phenyl-ethyl-, -propyl-, -butyl-, -hexyl-, -2-methylethyl-,-2-ethylethyl-, -2,2-dimethylethyl group; an o-, m-, p-methyl, ethyl,propyl, isopropylbenzyl group; a 2′,3′-, 2′,4′-, 2′,5′-, 2′,6′-, 3′,4′-,3′,5′-dimethyl- or -diethylbenzyl group; a 2′-, 3′-, 4′-fluoro-,chloro-, bromo-, iodobenzyl group; a 2′,3′-, 2′,4′-, 2′,5′-, 2′,6′-,3′,4′-, 3′,5′-, difluoro, dichloro-, dibromo- or diiodobenzyl group or a2- or 3-naphthylmethyl group; a 2-phenylethyl-, 3-phenyl-propyl-,4-phenylbutyl-, or 5-phenylpentyl group.

[0027] A C₁-C₆ alkyl group: A straight or branched alkyl group with 1-6C atoms, such as a methyl, ethyl, n-propyl, iso-propyl, n-, iso-,tert-butyl, n-pentyl, 2,2-dimethylpropyl or 3-methylbutyl group.

[0028] A cycloalkyl radical: A cyclopropane, cyclobutane, cyclopentane,cyclohexane, cycloheptane, or decahydronaphthalene radical.

[0029] A cycloalkylalkyl radical: A cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl-methyl radical; a 1-cyclopropyl,1-cyclobutyl, 1-cyclopentyl, 1-cyclohexyl, 1-cycloheptyl-ethyl radical;a 2-cyclopropyl, 2-cyclobutyl, 2-cyclopentyl, 2-cyclohexyl, or2-cycloheptyl-ethyl radical.

[0030] A heterocyclic ring: An unsubstituted 2- or 3-thienyl group or a2- or 3-furyl group or a 3-methyl-, 3-ethyl-, 3-fluoro-, 3-chloro-,3-bromo-, 3-iodo-2-furyl- or -2-thienyl group; a 4-methyl-, 4-ethyl,4-fluoro-, 4-chloro-, 4-bromo-, 4-iodo-2-furyl or -2-thienyl group; a5-methyl-, 5-ethyl-, 5-fluoro-, 5-chloro-, 5-bromo-, 5-iodo-2-furyl- or-2-thienyl group; a 2-methyl, 2-ethyl, 2-fluoro-, 2-chloro-, 2-bromo-,2-iodo-3-furyl or -3-thienyl group; a 4-methyl-, 4-ethyl-, 4-fluoro-,4-chloro-, 4-bromo-, 4-iodo-3-furyl- or -3-thienyl group; a 5-methyl-,5-ethyl-, 5-fluoro-, 5-chloro-, 5-bromo-, 5-iodo-3-furyl- or -3-thienylgroup; an unsubstituted 2-, 3- or 4-pyridyl group or a 3-methyl-,3-ethyl-, 3-fluoro-, 3-chloro-, 3-bromo-, 3-iodo-2-pyridyl group; a4-methyl-, 4-ethyl-, 4-fluoro-, 4-chloro-, 5-bromo-, 4-iodo-2-pyridylgroup; a 5-methyl-, 5-ethyl-, 5-fluoro-, 5-chloro-, 5-bromo-,5-iodo-2-pyridyl group; a 2-methyl-, 2-ethyl-, 2-fluoro-, 2-chloro-,2-bromo-, 2-iodo-3-pyridyl group; a 4-methyl-, 4-ethyl-, 4-fluoro-,4-chloro-, 4-bromo-, 4-iodo-3-pyridyl group; a 5-methyl-, 5-ethyl-,5-fluoro-, 5-chloro-, 5-bromo-, 5-iodo-3-pyridyl group; a 2-, 4-, 5-,6-pyrimidinyl group; a 3-, 4-, 5-, 6-pyridazinyl group or a 2- or3-pyrazinyl group.

[0031] R2 means:

[0032] An alkyl group: A straight or branched alkyl group with 1-6 Catoms, such as a methyl, ethyl, n-propyl, iso-propyl, n-, iso-,tert-butyl, n-pentyl, 2,2-dimethylpropyl or 3-methylbutyl group. Ahydrogen atom is preferred.

[0033] An optionally substituted phenyl ring or naphthyl ring: a phenylgroup; an o-, m-, p-methyl, -ethyl, -propyl-, isopropylphenyl group; a2,3-, 2,4-, 2,5-, 2,6-, 3,4-, 3,5-dimethyl- or -diethylphenyl group; ano-, m-, p- fluoro-, chloro-, bromo-, or iodophenyl group; a 2,3-, 2,4-,2,5-, 2,6-, 3,4-, 3,5-difluoro-, dichloro-, dibromo- or diiodophenylgroup; an o-, m-, p-trihalomethylphenyl group; a 2,3-, 2,4-, 2,5-, 2,6-,3,4- or 3,5 di-trihalogen-phenyl group; an o-, m-, p-methoxy, -ethoxy,-propoxy, -isopropoxyphenyl group or a naphthyl group. A2,5-difluorophenyl group is preferred.

[0034] R3 and R4 mean:

[0035] An alkyl group: A straight or branched alkyl group with 1-6 Catoms, such as a methyl, ethyl, n-propyl, iso-propyl, n-, iso-,tert-butyl, n-pentyl, 2,2-dimethylpropyl- or 3-methylbutyl group. Ahydrogen atom is preferred.

[0036] R5 means:

[0037] An alkyl group: A straight or branched alkyl group with 1-6 Catoms, such as a methyl-, ethyl-, n-propyl-, iso-propyl-, n-, iso-,tert-butyl-, n-pentyl-, 2,2-dimethylpropyl- or 3-methylbutyl group. Ahydrogen atom is preferred.

[0038] R6 means:

[0039] An alkyl group: A straight or branched alkyl group with 1-6 Catoms, such as a methyl-, ethyl-, n-propyl-, iso-propyl-, n-, iso-,tert-butyl-, n-pentyl-, 2,2-dimethylpropyl- or 3-methylbutyl group. Amethyl group is preferred.

[0040] An aralkyl group with 7-20 C atoms: A benzyl group; a1-phenyl-ethyl-, -propyl-, -butyl-, -hexyl-, -2-methylethyl-,-2-ethylethyl-, or -2,2-dimethylethyl group; an o-, m-, p-methyl-,ethyl-, propyl-, or isopropylbenzyl group; a 2′,3′-, 2′,4′-, 2′,5′-,2′,6′-, 3′,4′-, 3′,540 -dimethyl- or -diethylbenzyl group; a 2′-, 3′-,4′-fluoro-, chloro-, bromo-, or iodobenzyl group; a 2′,3′-, 2′,4′-,2′,5′-, 2′,6′-, 3′,4′-, 3′,5′-, difluoro-, dichloro-, dibromo- ordiiodobenzyl group or a 2- or 3-naphthylmethyl group; a 2-phenylethyl-,3-phenylpropyl-, 4-phenylbutyl, or 5-phenylpentyl group.

[0041] A heteroaralkyl group with 7-20 C atoms: A 2-, 3- or4-pyridyl-methyl-, -ethyl- or -propyl group; a 2- or 3-furyl-methyl-,-ethyl-, or -propyl group; a 2- or 3-thienyl-methyl-, -ethyl- or -propylgroup; a 2-, 3-, 4-, 5-, 6-, or 7-indolyl-methyl-, -ethyl- or -propylgroup. The benzyl group is preferred.

[0042] Preferred are compounds of formula (I), in which

[0043] —W═X═Y— is the group

[0044] If R1 is group —CO—R11, then R11 has, for example, the preferredmeaning of methyl, ethyl, i-propyl, phenyl, 2-thienyl and 2-furyl. If RIhas the meaning of —CO—OR12, then R12 can be, for example, preferablymethyl, ethyl or i-propyl.

[0045] In addition, compounds are preferred in which R2 is an aromaticgroup, e.g., a benzyl group, for example a 2′,6′-difluorobenzyl group,that is substituted on the aromatic ring by one or more halogen atoms,especially fluorine atoms. Also preferred are compounds in which atleast one of R³ and R⁴, especially both, are hydrogen atoms.

[0046] A. preferred meaning of Z is a direct bond or an oxygen atom,while G preferably means a —C═C group. L is preferably an NH group,while Q preferably is a carbonyl group and R5 1 is a C₁-C₆ allyl group.Especially preferred meanings of R61 are hydrogen atoms or C₁-C₃ alkylgroups, especially methyl groups, and an especially preferred meaning ofR62 is an aralkyl radical, e.g., a benzyl group.

[0047] The production of compounds (1) is preferably carried out

[0048] (a) By reaction of a compound of general formula (2)

[0049]  whereby R7 means a leaving group, e.g., a halogen atom or analkyl, perfluoroalkyl or arylsulfonyl group, and all other radicals havethe meaning that is indicated in compound (1), with a compound ofgeneral formula (3)

R8-N(R61)R62  (3)

[0050]  whereby R8 means a hydrogen atom or a metal atom, such as, e.g.,a lithium, sodium, potassium, cesium, calcium or barium atom, and R61and R62 have the meanings that are indicated in compound (1),

[0051] (b) By reaction of a compound of general formula (4)

[0052]  in which R9 is the group —OSO₂C_(n)F_(2n+1), a halogen atom,especially a bromine or iodine atom, or another leaving group, and allother radicals have the meaning that is indicated in compound (1), witha compound of general formula (5)

[0053]  whereby R10 is a group that contains a metal, such as a groupthat contains a trialkyltin group, a halomagnesium group or a group thatcontains a non-metal, such as boron, silicon, etc.; a dialkoxyborongroup or a dihydroxyboron group; a hydroxy or mercapto group that isoptionally converted into a metal salt, such as, e.g., a lithium,sodium, potassium, cesium, calcium, barium, silver or copper salt; thegroup —C≡C—R31 or an E- or Z-configured group —CR52=CR53R31 or—CR31=CR52R53, in which R31 is a group that contains a metal or anon-metal, such as boron, silicon, etc., such as a trialkyltin group, ahalomagnesium group, a dialkoxyboron group or a dihydroxyboron group,and all other radicals have the meaning that is indicated in compound(1), with or without the involvement of a catalyst, such as, e.g.,copper, nickel, palladium, platinum or organic derivatives of theabove-mentioned metals;

[0054] (c) If Y is a nitrogen atom in compound (1), by reaction of acompound of general formula (6)

[0055]  whereby R32 means a hydrogen atom or a metal atom, such as,e.g., a lithium, sodium, potassium, cesium, calcium, barium, silver orcopper atom, and all other radicals have the meaning that is indicatedin compound (1), with a compound of general formula (7)

R33-R2  (7)

[0056]  whereby R33 means a leaving group, e.g., a halogen atom or analkyl, perfluoroalkyl or arylsulfonyl group, and R2 has the meaning thatis indicated in compound (1) or

[0057] (d) If W in compound (1) is a nitrogen atom, by reaction of acompound of general formula (8)

[0058]  whereby R32 means a hydrogen atom or a metal atom, such as,e.g., a lithium, potassium, cesium, calcium, barium, silver or copperatom, and all other radicals have the meaning that is indicated incompound (1), with a compound of general formula (9)

R33-R1  (9)

[0059]  whereby R33 means a leaving group, e.g., a halogen atom or analkyl, perfluoroalkyl or arylsulfonyl group, and R1 has the meaning thatis indicated in compound (1).

[0060] Compounds (1) according to the invention can be used asantagonists of the gonadotropin-releasing hormone, for example for malebirth control, for hormone therapy, for treatment of female subfertilityand infertility, for female contraception and to combat tumors.

[0061] In male birth control, the compounds according to the inventionbring about a reduction in spermatogenesis. A combined administrationwith androgens, e.g., testosterone or testosterone derivatives, such as,for example, testosterone esters, preferably takes place. Theadministration of testosterone derivatives can be carried out, forexample, by injection, e.g., by intramuscular depot injection.

[0062] Compounds (1), optionally in combination with other hormones,e.g., estrogens and/or progestins, can also be used in hormone therapy,for example for treating endometriosis, uterus leiomyomas and uterinefibroids. Especially preferred are combinations of the GnRH antagonistsaccording to the invention and tissue-selective partial estrogenagonists such as Raloxifene(®). Moreover, compounds (1) according to theinvention can be used for increasing female fertility, for example byinducing ovulation, and treating sterility.

[0063] In contrast, compounds (1) are also suitable for contraception infemales. Thus, the GnRH antagonist can be administered on days 1 to 15of the cycle together with estrogen, preferably with very low estrogendosages. On days 16 to 21 of the intake cycle, progestagen is added tothe estrogen-GnRH-antagonist combination. The GnRH antagonist can beadministered continuously over the entire cycle time. In this way, areduction in the hormone dosages and thus a reduction in the sideeffects of unphysiological hormone levels can be achieved. In addition,advantageous effects in women who suffer from polycystic ovariansyndrome and androgen-dependent diseases, such as acne, seborrhea andhirsutism, can be achieved. An improved cycle monitoring relative toprevious administration methods can also be expected. Furtherindications are benign prostate hyperplasia, gonad protection inchemotherapy, controlled ovarian stimulation/artificial reproductiontechniques, and infantile development disorders, e.g., Pubertas praecoxand polycystic ovaries.

[0064] Finally, the GnRH agonists according to the invention can also beused for the treatment of hormone-dependent tumor diseases, such aspremenopausal breast cancer, prostate cancer, ovarian cancer andendometrial cancer, by the endogenous sex steroid hormones beingsuppressed.

[0065] Compounds (1) according to the invention are suitable as GnRHantagonists for administration to humans, but also for the purposes ofveterinary medicine, e.g., in the case of domestic and working animalsbut also in the case of wild animals.

[0066] The administration can be carried out in the known way, forexample, orally, topically, rectally, intravaginally, nasally or byinjections. Oral administration is preferred. Compounds (1) are broughtinto a form that can be administered and are optionally mixed withpharmaceutically acceptable vehicles or diluents. The oraladministration can be carried out, for example, in solid form astablets, capsules, coated tablets or powders, but also in the form of adrinkable solution. The non-oral administration can be carried out by,for example, intravenous, subcutaneous or intramuscular injection or byointments, creams or suppositories. An administration as a timed-releaseform can optionally also be carried out. The dosage can vary dependingon the type of indication, the severity of the disease, the age, sex,body weight and sensitivity of the subject to be treated. Dosages of0.01 to 30 mg, especially preferably 0.1 to 3 mg, and most preferably0.1 to 1 mg per kg of body weight and per day are preferablyadministered. The administration can be carried out in an individualdose or several separate dosages.

[0067] Below, a number of especially preferred compounds (1) are listed:Especially preferred compounds

6-(4-Acetamidophenyl)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylic acid-2-propyl ester

6-(4-Acetamidophenyl)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylic acid ethyl ester

6-(4-Acetamidophenyl)-3-isobutyryl-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline

6-(2-Acetamido-5-pyridyl)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluoro-benzyl)-1,4-dihydro-4-oxo-quinoline-3- carboxylic acid-isopropyl ester

6-(3-Acetamido-6-pyridyl)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluoro-benzyl)-1,4-dihydro-4-oxo-quinoline-3- carboxylic acid 2-propyl ester

6-(5-Acetamido-2-thienyl)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluoro-benzyl)-1,4-dihydro-4-oxo-quinoline-3- carboxylic acid-2-propyl ester

6-[1-(4-Acetamidophenyl)-vinyl]-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline- 3-carboxylic acid-ethylester

R- and S- and R,S-6-[1-(4-Acetamidophenyl)-ethyl]-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo- quinoline-3-carboxylicacid-ethyl ester

5-(N-Benzyl-N-methylamino-methyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-6-(4- methanesulfonylamidophenoxy)-4-oxo-quinoline-3-carboxylic acid-ethyl ester

6-(3-Acetamidophenoxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylic acid-ethyl ester

6-(2-Acetamido-5-pyridyloxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluoro-benzyl)-1,4-dihydro-4-oxo-quinoline-3- carboxylic acid-ethyl ester

6-(2-Methylaminocarbonyl-5-pyridyloxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline- 3-carboxylic acid-ethylester

1-[6-(4-Acetamidophenoxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluoro-benzyl)-1,4-dihydro-4-oxo-quinolin-3-yl)- methyl]-3-pyridin-2-yl-urea

1-[6-(4-Acetamidophenoxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluoro-benzyl)-1,4-dihydro-4-oxo-quinolin-3-yl)- oxy]-3-pyridin-2-yl-urea

6-(4-Hydroxyacetamidophenoxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline- 3-carboxylic acid-ethylester

6-(Acetamidophenoxy)-5-(N-benzyl-N- methylaminomethyl)-1-[bis-(2-fluoro-phenyl)-methyl]-1,4-dihydro-4-oxo- quinoline-3-carboxylic acid-ethylester

6-(Acetamidophenoxy)-5-(N-benzyl-N-methylaminomethyl)-1-diphenylmethyl-1,4-dihydro-4-oxo-quinoline-3-carboxylic acid- ethyl ester

[0068] In addition, the invention is to be explained by the followingexamples.

EMBODIMENTS EXAMPLE 16-(4-Acetamidophenoxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylic acid-ethylester

[0069]

[0070] 78 mg of 6-(4-acetamidophenoxy)-5-(chloromethyl)-1(2′,6′-difluorobenzyl) 1,4-dihydro-4-oxo-quinoline-3-carboxylicacid-ethyl ester, dissolved in 3 ml of dimethylformamide, was mixed with84 μl of N-methylbenzylamine and 84 μl of N,N-diisopropyl-ethylamine at0° C. and stirred at room temperature for 20 hours. After 20 ml ofaqueous sodium bicarbonate solution was added, the precipitate wassuctioned off, washed with water and then with n-hexane, and dried atroom temperature in a vacuum. 70 mg of the title compound is obtained.

[0071] NMR:=1.3 (t; 3H; CH₃); 1.9 (s; 3H; NCH₃); 2.05 (s; 3H; CH₃); 3.55(s; 2H; NCH₂); 4.27 (q; 2H; 0CH₂); 4.91 (s; 2H; NCH₂); 5.68 (s; 2H;NCH₂); 685 (d; 2H; ArCH); 7.1-7.22 (m; 7H; ArCH); 7.26 (d; 1H; ArCH);7.42-7.6 (m; 4H; ArCH); 8.72 (s; 1H; NCH); 9.9 (s; 1H; NH) MS: FAB:M^(⊕)+1=526 [M=525]

[0072] The starting material,6-(4-acetamidophenoxy)-5-(chloromethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid ethyl ester, was obtained in the following way:

[0073] a. 1,4-Dihydro-6-fluoro-5-nitro-4-oxo-quinoline-3-carboxylic acidethyl ester

[0074] 50 g of 4-fluoro-3-nitroaniline and 69 g of ethoxymethylenemalonic acid-diethyl ester were heated for 3 hours to 120° C. Themixture was added to n-hexane, stirred for 2 hours, and the crystallinematerial was suctioned off and dried in a vacuum at room temperature. 93g of N-(4-fluoro-3-nitrophenyl)-aminomethylene malonic acid diethylester is obtained. The latter is added in 3 portions of 31 g torespectively 150 ml of a mixture that consists of 26.5% diphenyl and73.5% diphenyl ether (DOWTHERM A^(⊕)) that is preheated to 260° C., andit is stirred for 30 minutes at this temperature. After cooling, it isdiluted with 500 ml of n-hexane, and the precipitate is suctioned off. Atotal of 64 g of a mixture of1,4-dihydro-6-fluoro-5-nitro-4-oxo-quinoline-3-carboxylic acid ethylester and 1,4-dihydro-6-fluoro-7-nitro-4-oxo-quinoline-3-carboxylicacid-ethyl ester is obtained.

[0075] NMR:=1.31 (t; 3H; CH₃); 4.25 (q; 2H; OCH₂); 7.88 (s; 1H; ArCH;isomer A); 7.92 (s; 1H; ArCH; isomer A); 8.1 (d; 1H; ArCH; isomer B);8.45 (d; 1H; ArCH; isomer B); 8.59+8.69 (2s; 1H each; NCH; A+B) MS: E I:M^(⊕)=280 [M=280]

[0076] b.1-(2′,6′-Difluorobenzyl)-1,4-dihydro-6-fluoro-5-nitro-4-oxo-quinoline-3-carboxylicacid ethyl ester

[0077] 22 g of the above-described mixture is stirred in 500 ml ofdimethylformamide with 16 g of potassium carbonate and 23.7 g of2,6-difluorobenzyl bromide for 5 hours at room temperature. The reactionmixture is added to 1 l of aqueous ammonium chloride solution andextracted three times with ethyl acetate. After drying with sodiumsulfate and concentration by evaporation in a vacuum, 500 ml of n-hexaneis added, and it is stirred for 15 minutes. After the hexane phase isdecanted, the residue is recrystallized from ethyl acetate. 10.4 g ofthe title compound is obtained.

[0078] NMR:=1.3 (t; 3H; CH₃); 4.25 (q; 2H; OCH₂); 5.84 (s; 2H; NCH₂);7.15-7.25 (m; 2H; ArCH); 7.45-7.55 (m; 1H; ArCH); 7.92 (dd; 1H; ArCH);8.04-8.14 (m; 1H; ArCH); 8.96 (s; 1H; NCH); MS: E I: M^(⊕)=406 [M=406]

[0079] c.6-(4-Acetamidophenoxy)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-5-nitro4-oxo-quinoline-3-carboxylicacid ethyl ester

[0080] 3.74 g of 4-acetamidophenol in 40 ml of dimethylformamide ismixed with 733 mg of sodium hydride (80% in mineral oil) and stirred for15 minutes at room temperature. Then, this solution is added to 5 g ofthe above-described compound, dissolved in 40 ml of dimethylformamide.After 5 hours at room temperature, the mixture is added to ice water,and the precipitate is suctioned off. After chromatography on silica gel(eluant dichloromethane/2-propanol 95:5), 5.17 g of the title compoundis obtained.

[0081] NMR:=1.3 (t; 3H; CH₃); 2.03 (s; 3H; CH₃); 4.27 (q; 2H; OCH₂);5.79 (s; 2H; NCH₂); 7.01 (d; 2H; ArCH); 7.13-7.23 (m; 2H; ArCH);7.43-7.56 (m; 2H; ArCH); 7.61 (d; 2H; ArCH); 7.8 (d; 1H; ArCH); 8.93 (s;1H; NCH); 9.97 (1H; S; NH) MS: E I: M^(⊕)=537 [M=537]

[0082] d.61-(4-Acetamidophenoxy)-5-amino-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid-ethyl ester

[0083] 5.1 g of the above-mentioned compound is hydrogenated in 650 mlof methanol with 510 mg of palladium/carbon (10%). After the catalyst issuctioned off and after concentration by evaporation, 4.55 g of thetitle compound is obtained.

[0084] NMR:=1.29 (t; 3H; CH₃); 2.02 (s; 3H; CH₃); 3.25 (s; 2H; NH₂);4.25 (q; 2H; OCH₂); 5.55 (s; 2H; NCH₂); 6.55 (d; 1H; ArCH); 6.86 (d; 2H;ArCH); 7.07 (d; 1H; ArCH); 7.1-7.22 (m; 2H; ArCH); 7.4-7.55 (m; 3H;ArCH); 8.71 (s; 1H; NCH); 9.8 (s; 1H, NH); MS: E I: M^(⊕)=507 [M=507]

[0085] e.5-(4-Acetamidophenoxy)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-5-iodo-4-oxo-quinoline-3-carboxylicacid-ethyl ester

[0086] 2 g of the above-mentioned compound is dissolved in a mixture of24 ml of concentrated sulfuric acid and 12 ml of water, and it is mixedat 0° C. with 325 mg of sodium nitrite in 4 ml of water. After 15minutes, a pH of 3 is set with sodium bicarbonate solution, and 100 mgof urea is added. Then, 723 mg of potassium iodide in 0.5 ml of water isadded, and it is stirred for one hour at room temperature. Afterextraction with dichloromethane/methanol (95:5, v/v), the organic phaseis washed with aqueous sodium thiosulfate solution, dried andconcentrated by evaporation. After chromatography on silica gel (eluantdichloromethane with 0-15% isopropanol), 697 mg of the title compound isobtained.

[0087] NMR:=1.3 (t; 3H; CH₃); 2.02 (s; 3H; CH₃); 4.25 (q; 2H; OCH₂);5.71 (s; 2H; NCH₂); 6.83 (d; 2H; ArCH); 7.1-7.2 (2H; m; ArCH); 7.3 (d;1H; ArCH); 7.41-7.63 (m; 4H; ArCH); 8.83 (s; 1H; N—CH); 9.95 (s; 1H; NH)MS: es: M^(⊕)+1=493 [M=492]

[0088] f.6-i(4-Acetamidophenoxy)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-5-styryl-quinoline-3-carboxylicacid-ethyl ester

[0089] 5:50 mg of the above-mentioned iodine compound, 198 mg ofstyrylboronic acid, 55 mg of tetrakis-triphenylphosphine-palladium(O),1.1 ml of 2 molar sodium carbonate solution, 2.2 ml of ethanol and 22 mlof toluene are stirred for 6 hours at 80° C. Then, water is added,extracted with dichloromethane, and the organic phase is washed withcommon salt solution, dried and concentrated by evaporation. The residueis purified by chromatography on silica gel (eluant dichoromethane with0-10% isopropanol). 902 mg of the title compound is obtained.

[0090] EMR:=1.28 (t; 3H; CH₃); 2.0 (s; 3H; CH₃); 4.24 (q; 2H; OCH₂); 5.7(s; 2H; N—CH₂); 6.75-6.85 (m; 3H; ArCH; CH═CH); 7.08-7.28 (m; 3H; ArCH);7.3-7.68 (m; 9H; ArCH); 7.86 (d; 1H; CH═CH); 8.75 (S; 1H; NCH); 9.72(1H; S; NH) MS: es: M^(⊕)+1=595 [M=594]

[0091] g.6(4-Acetamidophenoxy)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-5-formyl-4-oxo-quinoline-3-carboxylicacid-ethyl ester

[0092] 450 mg of the above-mentioned styryl compound is dissolved in 25ml of tetrahydrofuran and 7 ml of water, and it is mixed with 0.11 ml ofa 2.5% solution of osmium tetroxide in tert-butanol. After 15 minutes ofstirring at room temperature, 482 mg of sodium periodate is added; after20 and 22 hours, 100 mg of sodium periodate is added; and after 24hours, another 100 mg of sodium periodate is added. After 26 hours, itis diluted with water and extracted with ethyl acetate. After theorganic phase is dried with sodium sulfate, it is concentrated byevaporation. 352 mg of the title compound is obtained as a foam.

[0093] NMR:=1.29 (t; 3H; CH₃); 2.01 (s; 3H; CH₃); 4.27 (q; 2H; OCH₂);5.71 (s; NCH₂); 6.91 (d; 2H; ArCH); 7.12-7.22 (m; 2H; ArCH); 7.4 (d; 1H;ArCH); 7.43-7.64 (m; 3H; ArCH); 7.72 (d; 1H; ArCH); 9.0 (d; 1H; NCH);9.97 (s; 1H; NH); 10.44 (s; 1H; CHO) MS: FAB: M^(⊕)+1=521 [M=520]

[0094] h.6-(4-Acetamidophenoxy)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-5-hydroxymethyl-4-oxo-quinoline-3-carboxylicacid-ethyl ester

[0095] 300 mg of the above-mentioned aldehyde is dissolved in 13.8 ml ofacetic acid and mixed at 10-minute intervals with 3 portions of 10 mg ofsodium borohydride each. After dilution with water, it is extracted withethyl acetate, and the organic phase is washed neutral with sodiumbicarbonate solution. After drying with sodium sulfate, it isconcentrated by evaporation. By chromatography on silica gel (eluantdichloromethane with 0-10% isopropanol), 129 mg of the title compound isobtained as a foam.

[0096] MNR:=1.31 (t; 3H; CH₃); 2.02 (s; 3H; CH₃); 4.27 (q; 2H; OCH₂);4.86 (d; 2H; OCH2); 5.12 (t; 1H; OH); 5.8 (s; 2H; NCH₂); 6.85 (d; 2H;ArCH); 7.12-7.22 (m; 2H; ArCH); 7.38 (d; 1H; ArCH); 7.42-7.59 (m; 3H;ArCH); 7.63 (d; 1H; ArCH); 8.94 (s; 1H; NCH); 9.5 (s; 1H, NH) MS: FAB:M^(⊕)+1=523 [M=522]

[0097] i. 6-(4-Acetamidophenoxy)-5-chloromethyl-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-(quinoline-3-carboxylicacid-ethyl ester

[0098] 80 mg of the above-mentioned alcohol is dissolved in 2 ml ofdichloromethane and mixed with 0. 1 ml of thionyl chloride. After 20minutes at room temperature, water is added, it is extracted withdichloromethane, and the organic phase is washed with common saltsolution. After drying with sodium sulfate, it is concentrated byevaporation. 80 mg of the title compound is obtained as a foam.

[0099] NMR:=1.33 (t; 3H; CH₃); 2.07 (s; 3H; CH₃); 4.3 (q; 2H; OCH₂);5.66 (s; 2H; CH₂Cl); 5.75 (s; 2H; NCH₂); 6.98 (d; 2H; ArCH); 7.13-7.25(m; 2H; ArCH); 7.3 (d; 1H; ArCH); 7.53 (t; 1H; ArCH); 7.61 (d; 2H;ArCH); 7.68 (d; 1H; ArCH); 8.82 (s; 1H; NCH); 9.98 (s; 1H; NH) MS: es:M^(⊕)+1=541/543 [M=540/542]

EXAMPLE 26-(4-Acetamidophenoxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluoro-benzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid-2-propyl ester

[0100]

[0101] This compound is produced from6-(4-acetamidophenoxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid ethyl ester by heating with titanium-tetraisopropylate inisopropanol.

EXAMPLE 35-(N-Benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-6-(4-isobutyramidophenoxy)-4-oxo-quinoline-3-carboxylicacid-ethyl ester

[0102]

[0103] This compound is produced analogously to Example 1 from5-(chloromethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-6-(4isobutyramidophenoxy)-4-oxo-quinoline-3-carboxylicacid-ethyl ester and N-methylbenzylamine as a foam.

[0104] a.5-(Chloromethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-6-(4-isobutyramidophenoxy)-4-oxo-quinoline-3-carboxylicacid-ethyl ester

[0105] This compound is obtained when, in Example 1/c,4-isobutyramidophenol is used instead of 4-acetamidophenol and the restof the reaction is carried out as described in Example 1/d. to Example1/i.

EXAMPLE 43-Acetyl-6-(4-acetamidophenyl)-1-benzyl-5-(N-benzyl-N-methylaminomethyl)-phthalazin-4-one

[0106]

[0107] The title compound is obtained by6-(4-acetamidophenyl)-1-benzyl-5-(N-benzyl-N-methylaminomethyl)-phthalazin-4-onebeing reacted in the presence of a base such as sodium carbonate orsodium hydroxide with acetyl chloride or acetic anhydride.

[0108]6-(4-Acetamidophenyl)-1benzyl-5-(N-benzyl-N-methylaminomethyl)-phthalazin-4-oneis obtained in the following way:

[0109] a. 6-(4-Acetamidophenyl)-1-benzyl-5-iodo-phthalazin4-one

[0110] 1-Benzyl-5,6-diiodo-phthalazin-4-one (Indian J. Chem. 16B, 1978,301-304) is reacted analogously to Example 1/f. with 1 equivalent of4-acetamidophenyl-boronic acid. The title compound is obtained in pureform by chromatography on silica gel.

[0111] b. 6-(4-Acetamidophenyl)-1-benzyl-5-chloromethyl-phthalazin-4-one

[0112] 6-(4-Acetamidophenyl)-1-benzyl-5iodo-phthalazin-4-one is furtherreacted to form the title compound analogously to Example 1/f.-i.

EXAMPLE 56-(4-Acetamidophenoxy)-5-(N-benzyl-N-methylamino-methyl)-1,4-dihydro-4-oxo-1-(2′-trifluoromethylbenzyl)-quinoline-3-carboxylicacid-ethyl ester

[0113]

[0114] 130 mg (0.226 mmol) of6-(4-acetamidophenoxy)-5-(chloromethyl)-1,4-dihydro-4-oxo-1-(2′-trifluoromethylbenzyl)-quinoline-3-carboxylicacid-ethyl ester, dissolved in 5 ml of DMF, is mixed at −5° C. with 125μl of N,N-diisopropyl-ethylamine and 126 μl (0.97 mmol) ofN-benzyl-methylamine. After heating to room temperature, it is allowedto stir for 20 more hours, and then the reaction mixture is added to 50ml of saturated sodium bicarbonate solution. The solid is suctioned off,washed with water and dried in a vacuum. The additional purification iscarried out by chromatography on silica gel with an eluant that consistsof 90 parts of dichloromethane, 10 parts of ethanol and 1 part ofconcentrated ammonia (R_(f): 0.38).2.

45 mg of the title compound is obtained as a foam. MS/molar peak, M⁺=658

[0115] The starting material6-(4-acetamidophenoxy)-5-(chloromethyl)-1,4-dihydro-4-oxo-1-(2′-trifluoromethylbenzyl)-quinoline-3-carboxylicacid-ethyl ester is produced analogously to the method that is describedin Examples 1a to 1i with use of 2′-trifluoromethyl-benzyl bromideinstead of 2,6-difluoromethylbenzyl bromide.

EXAMPLE 66-(4-Methylaminocarbonyl-phenoxy)-5-(N-benzyl-N-methylaminomethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid-ethyl ester

[0116] 35 mg (0.061 mmol) of6-(4-methylaminocarbonyl-phenoxy)-5-(chloromethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid-ethyl ester, dissolved in 1.3 μl of DMF, is mixed at 0° C. with 35μl of N,N-diisopropyl-ethylamine and 35 μl (0.25 mmol) ofN-methylbenzylamine. After heating to room temperature, it is allowed tostir for 20 more hours, and then the reaction mixture is added to 10 mlof saturated sodium bicarbonate solution. The accumulated solid issuctioned off, washed with water and hexane and dried on phosphoruspentoxide in a vacuum.

[0117] 27 mg of the title compound is obtained as a foam. MS (esi):M⁺+1=626 [M=625]

[0118] The starting material6-(4-methylaminocarbonyl-phenoxy)-5-(chloromethyl)-1-(2′,6′-difluorobenzyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid-ethyl ester is produced analogously to the method that is describedin Examples 1a to 1i with use of 4-hydroxy-N-methylbenzoic acid amideinstead of 4-acetamidophenol.

EXAMPLE 76-(4-Acetamidophenoxy)-5-(N-benzyl-N-methylamino-methyl)-1-(1-naphthyl-methyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid-ethyl ester

[0119] 45 mg (0.081 mmol) of6-(4-acetamidophenoxy)-5-(chloromethyl)-1-(1-naphthyl-methyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid-ethyl ester, dissolved in 1.7 ml of DMF, is mixed at 0° C. with 46μl of N,N-diisopropyl-ethylamine and 46 g (0.33 mmol) ofN-methylbenzylamine. After heating to room temperature, it is allowed tostir for 20 more hours, and then the reaction mixture is added to 15 mlof saturated sodium bicarbonate solution. The accumulated solid issuctioned off, washed with water and hexane and dried on phosphoruspentoxide in a vacuum.

[0120] 32 mg of the title compound is obtained as a foam. MS (esi):M⁺+1=639 [M=638]

[0121] The starting material6-(4-acetamidophenoxy)-5-(chloromethyl)-1-(1-naphthyl-methyl)-1,4-dihydro-4-oxo-quinoline-3-carboxylicacid-ethyl ester is produced analogously to the method that is describedin Examples 1a to 1i with use of 1-chloromethylnaphthalene instead of2,6-difluoromethylbenzyl bromide.

EXAMPLE 8 Detection of the Antagonistic Action

[0122] a) Materials

[0123] Buserelin was ordered from Welding (Frankfurt/Main, Germany). Thecompound was labeled with ¹²⁵I by use of the chloramine T-method andNa¹²⁵I(4000 Ci/mmol; Amersham-Buchler, Brunswick, Germany). The labeledsubstance was purified by reverse phase HPLC on a Spherisorb ODS IIcolumn (250×4 mm, particle size 3 μm) by elution with 50%acetonitrile/0.15% trifluoroacetic acid at a flow rate of 0.5 ml/min.The specific activity was 2000 Ci/mmol.

[0124] All other chemicals were ordered from commercial sources at thehighest available purity.

[0125] b) Cell culture

[0126] Alpha T3-1 cells (Bilezikjian et al., Mol. Endocrinol 5 (1991),347-355) were cultivated in DMEM medium (Gibco-BRL,Eggenstein-Leopoldshafen, Germany) with penicillin (100 I.U./ml),streptomycin (0.1 mg/ml) and glutamine (0.01 mol/l) and 10% fetal calfserum (FCS; PAA Laboratories, Coelbe, Germany) on plastic tissue cultureplates (Nunc, 245×245×20 mm). CHO-3 cells (Schmid et al., J. Biol. Chem.275 (2000), 9193-9200) were cultivated under identical conditions, apartfrom the fact that Ham's F12 medium (Gibco-BRL) was used.

[0127] Ten confluent cell culture plates were flushed twice with 50 mlof phosphate-buffered salt solution (PBS). The cells were harvested byscraping them off with a rubber scraper in 5 ml of PBS and sedimented bycentrifuging in a laboratory centrifuge (Heraeus) at 800 rpm for 10minutes. The cell pellet was resuspended in 5 ml of 0.25 mol/l ofsaccharose/0.01 mol/l of triethanolamine, pH 7.4. The cells were lysedby three cycles of freezing in dry ice/ethanol bath and thawing at roomtemperature. The lysate was centrifuged at 900 rpm for 10 minutes, andthe sediment was discarded. The supernatant was centrifuged at 18,000rpm in a Sorvall SS34 rotor for 30 minutes. The pellet (cell membranes)was suspended by Potters in 5 ml of assay buffer (0.25 mol/l ofsaccharose, 0.01 mol/l of triethanolamine, pH 7.5, 1 mg/ml of ovalbumin)and stored in 200 μl of aliquots at −20° C. The determination of proteinwas carried out according to the Bradford method (Anal. Biochem. 72(1976), 248-254).

[0128] c. Receptor assay

[0129] Binding studies for competition curves were performed astriplicates. A test sample contained 60 μl of cell membrane suspension(10 μg of protein for aT3-1 cells or 40 μg of protein for CHO3 cells),20 μl of ¹²⁵I-labeled buserelin (100,000 Ipm per sample for competitioncurves and between 1,500 and 200,000 Ipm for saturation experiments) and20 μl of test buffer or test compound solution. The test compounds weredissolved in distilled water or 50% ethanol. Serial dilutions (5×10⁻⁶mol/l to 5×10⁻¹² mol/l) were produced in test buffer. The unspecificbinding was determined in the presence of excess unlabeled buserelin(10⁻⁶ mol/l). The test samples were incubated for 30 minutes at roomtemperature. Bonded and free ligands were separated by filtration(Whatman GF/C-filter, 2.5 cm diameter) with use of an Amicon10×collecting device and washed twice with 5 ml of 0.02 mol/l Tris/HCl,pH 7.4. The filters were moistened with 0.3% polyethylenimine (Serva;Heidelberg, Germany) for 30 minutes to reduce the unspecific binding.The radioactivity that was held up by the filter was determined in a5-channel gamma-counter (Wallac-LKB 1470 Wizard).

1. Compounds of general formula (I):

in which R¹ (a) is an acyl group —CO—R11 or CN, whereby R11 is asaturated, unsaturated, cyclic and/or (hetero)aromatic organic radical,especially a straight or branched alkyl chain with 1-10 C atoms or aphenyl, furan or thiophene group that is optionally substituted by alkylgroups or halogen atoms, (b) is a carboxylic acid ester group —CO—OR12or a carboxylic acid amide group —CO—NR12R13 or a group —SO_(x)—R12 withX=0, 1 or 2 or —SO₂—NR12R13, whereby R12 is a saturated, unsaturated,cyclic and/or (hetero)aromatic organic radical, especially a straight orbranched alkyl chain with 1-10 C atoms, an aralkyl group with 7-20 Catoms, whereby the aryl radical optionally can be substituted by alkylgroups or halogen atoms or is a phenyl radical that is optionallysubstituted by alkyl groups or halogen atoms, and R13 can be a hydrogenatom or a straight or branched alkyl chain with 1-10 C atoms, or (c) isthe group —A—NR14—CO—NR15R16, in which A is an alkylene group with 1-4 Catoms, especially with 1 C atom, that is optionally substituted by aC₁-C₆ alkyl group, a carbonyl group, an oxygen atom or the group—SO_(x)— with X=0, 1 or 2; R14 and R15, in each case independently are ahydrogen atom or a straight or branched alkyl chain with 1-10 C atoms,and R16 is a straight or branched alkyl chain with 1-10 C atoms, acycloalkyl group with 3-10 C atoms, a cycloalkylalkyl group with 7-20 Catoms, an aralkyl group with 7-20 C atoms, whereby the aryl radicaloptionally can be substituted by alkyl groups or halogen atoms, a phenylgroup that is optionally substituted by alkyl groups or halogen atoms ora heterocyclic ring that is optionally substituted by alkyl groups orhalogen atoms, R2 is a group —CH(R21)R22, whereby R21 is a hydrogenatom, a C₁-C₁₀-alkyl group or an optionally substituted phenyl ring andR22 is an optionally substituted phenyl ring or naphthyl ring, or agroup —CH₂CH(R23)R24, with R23 and R24 in the meaning of an optionallysubstituted phenyl ring, R3 and R4 in each case independently are ahydrogen atom or an alkyl group with 1-10 C atoms and R3 also can be ahalogen atom, R5 is a group that is linked via radical Z,

 in which G is —C═C—, —C═N—, —N═C—, an oxygen or sulfur atom, Z is adirect bond, an oxygen atom or a sulfur atom, the group CH—52 or—CHR52—CH—R53-, whereby R52 and R53, independently of one another, havethe meaning of a hydrogen atom or an alkyl group and n means numbers 1and 2, a —C≡C-triple bond or an E- or Z-configured group —CR52═CR53- orC═CR52R53, whereby R52 and R53, independently of one another, have themeaning of a hydrogen atom or an alkyl group, L is a CH₂ group or an NHgroup, Q is a carbonyl group or —SO_(x) group, with X=0, 1 or 2, and R51is an amino group that is optionally substituted by an alkyl group or astraight or branched alkyl group that is optionally substituted byhalogen atoms, hydroxyl or alkoxy groups, or a cycloalkyl group with 3-7ring members that is optionally substituted by halogen atoms, hydroxylor alkoxy groups, R6 is the group CH₂—N(R61)R62, whereby R61, in eachcase independently, is a hydrogen atom or an alkyl group, and R62 is analkyl group or an optionally substituted aralkyl group or aheteroarylalkyl group with 7-20 C atoms, and can mean —W═X═Y— the groups

 in any orientation; also all stereoisomers of the above-mentionedstructures and salts thereof with physiologically compatible acids orbases.
 2. Compounds according to claim 1, characterized in that —W-X-Y-is the group


3. Compounds according to claim 1 or 2, wherein R1 is the group —CO—R11.4. Compounds according to claim 3, wherein R11 is selected from methyl,ethyl, i-propyl, phenyl, 2-thienyl and 2-furyl.
 5. Compounds accordingto claim 1 or 2, wherein R1 is the group —CO—OR12.
 6. Compoundsaccording to claim 5, wherein R12 is selected from methyl, ethyl ori-propyl.
 7. Compounds according to one of claims 1 to 6, wherein R2 isa 2′,5′-difluorobenzyl group.
 8. Compounds according to one of claims 1to 7, wherein R3 and R4 are hydrogen atoms.
 9. Compounds according toone of claims 1 to 8, wherein Z is a direct bond or an oxygen atom. 10.Compounds according to one of claims 1 to 9, wherein G—C═C—. 11.Compounds according to one of claims 1 to 10, wherein L is an NH group.12. Compounds according to one of claims 1 to 11, wherein Q is acarbonyl group, and R51 is a C₁-C₆ alkyl group.
 13. Compounds accordingto one of claims 1 to 12, wherein R61 is a hydrogen atom or a methylgroup and/or R62 is a benzyl group.
 14. Use of compounds according toone of claims 1 to 13 as antagonists of the gonadotropin-releasinghormone (GnRH).
 15. Use according to claim 14 for male birth control,for hormone therapy, for treating female subfertility and infertility,for female contraception and to combat tumors.
 16. Process for theproduction of compounds of general formula (1) (a) By reaction of acompound of general formula (2)

 whereby R7 means a leaving group, and all other radicals have themeaning that is indicated in compound (1), with a compound of generalformula (3) R8-N(R61)R62  (3)  whereby R8 means a hydrogen atom or ametal atom, and R61 and R62 have the meanings that are indicated incompound (1), (b) By reaction of a compound of general formula (4)

 in which R9 is the group —OSO₂CnF_(2n+1), a halogen atom, especially abromine or iodine atom, or another leaving group, and all other radicalshave the meaning that is indicated in compound (1), with a compound ofgeneral formula (5)

 whereby R10 is a group that contains a metal or a non-metal, a hydroxyor mercapto group that is optionally converted into a metal salt; thegroup —C≡C-R31 or an E- or Z-configured group —CR52=CR53R31 or—CR31=CR52R53, in which R31 is a group that contains a metal or anon-metal, and all other radicals have the meaning that is indicated incompound (1), with or without the involvement of a catalyst; (c) If Y isa nitrogen atom in compound (1), by reaction of a compound of generalformula (6)

 whereby R32 means a hydrogen atom or a metal atom, and all otherradicals have the meaning that is indicated in compound (1), with acompound of general formula (7) R33-R2  (7)  whereby R33 means a leavinggroup, and R2 has the meaning that is indicated in compound (1), or (d)If W in compound (1) is a nitrogen atom, by reaction of a compound ofgeneral formula (8)

 whereby R32 means a hydrogen atom or a metal atom, and all otherradicals have the meaning that is indicated in compound (1), with acompound of general formula (9) R33-R1  (9)  whereby R33 means a leavinggroup, and R1 has the meaning that is indicated in compound (1).